The auxin-inducible degradation system enables conditional PERIOD protein depletion in the nervous system of Drosophila melanogaster.
The auxin-inducible degradation system enables conditional PERIOD protein depletion in the nervous system of Drosophila melanogaster.
复制标题
生长素诱导的降解系统能够在黑腹果蝇的神经系统中条件性地消耗 PERIOD 蛋白。
DOI：
10.1111/febs.14677
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发表时间：
2018-12
期刊：
影响因子：
--
通讯作者：
Zhang Y
中科院分区：
文献类型：
--
作者：
Chen W;Werdann M;Zhang Y
能够诱导和可逆地消耗靶蛋白的工具对于生物学研究至关重要。植物来源的生长素诱导降解系统 (AID) 能够降解标有 AID 基序的目标蛋白。该系统最近已应用于哺乳动物细胞以及线虫和果蝇。为了测试 AID 方法在神经系统中的实用性，我们使用昼夜运动节律作为模型，并应用 AID 方法在时间和空间上降解 PERIOD（PER），这是果蝇中的一种关键起搏器蛋白。我们发现通过基于 CRISPR/Cas9 的基因组编辑可以有效地用 AID 基序标记周期基因座，而不会破坏 PER 功能。此外，我们证明 AID 系统可用于诱导神经系统中快速有效的蛋白质降解，如对昼夜节律和睡眠行为的影响所示。此外，去除生长素后，AID 引起的蛋白质降解可快速逆转。总之，我们的结果表明 AID 系统为果蝇行为研究提供了强大的工具。能够在时间和空间上快速消耗目标蛋白的工具对于生物学研究至关重要。最近开发了生长素诱导降解系统（AID），能够降解标有 AID 基序的靶蛋白。在这里，我们使用果蝇昼夜节律作为原理证明来测试 AID 系统在神经系统中的潜在用途。我们报告，AID 系统可用于引发果蝇神经系统中 PERIOD 蛋白的快速和可逆降解。
Tools that allow inducible and reversible depletion of target proteins are critical for biological studies. The plant-derived auxin-inducible degradation system (AID) enables the degradation of target proteins tagged with the AID motif. This system has been recently employed in mammalian cells as well as in C. elegans and Drosophila. To test the utility of the AID approach in the nervous system, we used circadian locomotor rhythms as a model and applied the AID method to temporally and spatially degrade PERIOD (PER), a critical pacemaker protein in Drosophila. We found that the period locus can be efficiently tagged with the AID motif by CRISPR/Cas9-based genome editing without disrupting PER function. Moreover, we demonstrated that the AID system could be used to induce rapid and efficient protein degradation in the nervous system as shown by effects on circadian and sleep behaviors. Furthermore, the protein degradation by AID was rapidly reversible after auxin removal. Together, our results show that the AID system provides a powerful tool for behavior studies in Drosophila. Tools that allow rapid temporal and spatial depletion of target proteins are critical for biological studies. The auxin-inducible degradation system (AID) has been recently developed to enable the degradation of target proteins tagged with the AID motif. Here we used the Drosophila circadian locomotor rhythms as a proof of principle to test the potential use of the AID system in the nervous system. We report that the AID system can be used to elicit rapid and reversible degradation of PERIOD protein in the Drosophila nervous system.
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